
A method, called "protein blotting," for the detection of DNA-binding proteins is described. Proteins are separated on an SDA-polyacrylamide gel. The gel is sandwiched between 2 nitrocellulose filters and the proteins allowed to diffuse out of the gel and onto the filters. The proteins are tightly bound to each filter, producing a replica of the original gel pattern. The replica is used to detect DNA-binding proteins, RNA-binding proteins or histone-binding proteins by incubation of the filter with [32P]DNA, [125I]RNA, or [125I] histone. Evidence is also presented that specific protein-DNA interactions may be detected by this technique; under appropriate conditions, the lac repressor binds only to DNA containing the lac operator. Strategies for the detection of specific protein-DNA interactions are discussed.
DNA Helicases, Chromatography, Gel / instrumentation, HeLa Cells / analysis, Histones, Carrier Proteins / analysis, Nucleoproteins, Electrophoresis, Polyacrylamide Gel / methods, Chromatography, Gel, DNA Helicases / analysis, Humans, RNA, Viral, Electrophoresis, Polyacrylamide Gel, Chromatography, Gel / methods, Nucleoproteins / analysis, Carrier Proteins, HeLa Cells
DNA Helicases, Chromatography, Gel / instrumentation, HeLa Cells / analysis, Histones, Carrier Proteins / analysis, Nucleoproteins, Electrophoresis, Polyacrylamide Gel / methods, Chromatography, Gel, DNA Helicases / analysis, Humans, RNA, Viral, Electrophoresis, Polyacrylamide Gel, Chromatography, Gel / methods, Nucleoproteins / analysis, Carrier Proteins, HeLa Cells
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